Altered vascular function after adenovirus-mediated overexpression of endothelial nitric oxide synthase

1997 ◽  
Vol 273 (1) ◽  
pp. H265-H270 ◽  
Author(s):  
H. Ooboshi ◽  
Y. Chu ◽  
C. D. Rios ◽  
F. M. Faraci ◽  
B. L. Davidson ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Endothelial Nitric Oxide Synthase ◽  
Vascular Function ◽  
Transgene Expression ◽  
A 23187 ◽  
Adventitial Cells ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide ◽  
Beta Galactosidase

Gene transfer with replication-deficient adenovirus is a potentially useful tool to study vascular biology. We have constructed a replication-deficient adenovirus (AdRSVeNOS) that carries cDNA for endothelial nitric oxide synthase (eNOS). Transfection of COS-1 cells with AdRSVeNOS increased nitric oxide synthase activity (measured as production of L-citrulline from L-arginine) that was calcium dependent and inhibited by N omega-nitro-L-arginine methyl ester. To investigate effects of overexpression of eNOS on vascular function, we incubated common carotid arteries from rabbits in organ culture with AdRSVeNOS or AdRSV beta gal encoding beta-galactosidase. Transgene expression and responses to vasoactive agents were examined 1 day after transduction. Histochemical staining of beta-galactosidase and immunohistochemistry for eNOS indicated transgene expression in endothelium and adventitial cells. After precontraction with phenylephrine, vessels treated with AdRSVeNOS demonstrated greater relaxation to acetylcholine than vessels treated with vehicle or AdRSV beta gal. Relaxation to calcium ionophore A-23187 was much greater in vessels treated with AdRSVeNOS than in vessels treated with vehicle or AdRSV beta gal. Augmented relaxation in response to A-23187 was also observed after denudation of endothelium in vessels treated with AdRSVeNOS and was inhibited by N omega-nitro-L-arginine. Thus vasorelaxation in response to stimuli that release nitric oxide is augmented after adenovirus-mediated overexpression of eNOS. Transgene expression in adventitial cells appears to be sufficient to alter vasomotor function.

scholarly journals Gene transfer of endothelial nitric oxide synthase (eNOS) in eNOS-deficient mice

1999 ◽  
Vol 277 (2) ◽  
pp. H770-H776 ◽  
Author(s):  
Kristy D. Lake-Bruse ◽  
Frank M. Faraci ◽  
Edward G. Shesely ◽  
Nobuyo Maeda ◽  
Curt D. Sigmund ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Gene Transfer ◽  
Endothelial Nitric Oxide Synthase ◽  
Vascular Function ◽  
Calcium Ionophore ◽  
A 23187 ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide

Relaxation to acetylcholine (ACh) and calcium ionophore (A-23187) is absent in aortas from endothelial nitric oxide synthase (eNOS)-deficient (eNOS -/-) mice. We hypothesized that gene transfer of eNOS would restore relaxation to ACh and A-23187 in eNOS -/- mice. Aortic rings from eNOS -/- and eNOS +/+ mice were exposed in vitro to vehicle or adenoviral vectors encoding β-galactosidase (lacZ) or eNOS. Histochemical staining for β-galactosidase and eNOS demonstrated transduction of endothelial cells and adventitia. Vehicle-treated vessels from eNOS -/- mice did not relax to ACh or A-23187 compared with eNOS +/+ mice. In contrast, relaxation to nitroprusside (NP) was significantly greater in eNOS -/- mice than in eNOS +/+ mice. Gene transfer of eNOS, but not lacZ, to vascular rings of eNOS -/- mice restored relaxation to ACh and A-23187. In vessels from eNOS -/- mice that were transduced with eNOS, N ω-nitro-l-arginine (10−4 M) inhibited relaxation to ACh and A-23187 but not NP. Thus vascular function can be significantly improved by gene transfer in vessels where a major relaxation mechanism is genetically absent.

Effect of a previous pregnancy on vascular function in endothelial nitric oxide synthase 3 knockout mice

2007 ◽  
Vol 197 (3) ◽  
pp. 279.e1-279.e5 ◽  
Author(s):  
Labib M. Ghulmiyyah ◽  
Esther Tamayo ◽  
Shannon M. Clark ◽  
Gary D.V. Hankins ◽  
Garland D. Anderson ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Knockout Mice ◽  
Endothelial Nitric Oxide Synthase ◽  
Vascular Function ◽  
Previous Pregnancy ◽  
Nitric Oxide Synthase 3 ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide

scholarly journals Endothelial nitric oxide synthase activation and nitric oxide function: new light through old windows

2011 ◽  
Vol 210 (3) ◽  
pp. 239-241 ◽  
Author(s):  
Ian M Bird
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Endothelial Nitric Oxide Synthase ◽  
Vascular Function ◽  
Calmodulin Binding ◽  
And Function ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide ◽  
First Time ◽  
New Evidence

The principle mechanisms operating at the level of endothelial nitric oxide synthase (eNOS) itself to control its activity are phosphorylation, the auto-regulatory properties of the protein itself, and Ca2+/calmodulin binding. It is now clear that activation of eNOS is greatest when phosphorylation of certain serine and threonine residues is accompanied by elevation of cytosolic [Ca2+]i. While eNOS also contains an autoinhibitory loop, Rafikov et al. (2011) present the evidence for a newly identified ‘flexible arm’ that operates in response to redox state. Boeldt et al. (2011) also review the evidence that changes in the nature of endothelial Ca2+ signaling itself in different physiologic states can extend both the amplitude and duration of NO output, and a failure to change these responses in pregnancy is associated with preeclampsia. The change in Ca2+ signaling is mediated through altering capacitative entry mechanisms inherent in the cell, and so many agonist responses using this mechanism are altered. The term ‘adaptive cell signaling’ is also introduced for the first time to describe this phenomenon. Finally NO is classically regarded as a regulator of vascular function, but NO has other actions. One proposed role is regulation of steroid biosynthesis but the physiologic relevance was unclear. Ducsay & Myers (2011) now present new evidence that NO may provide the adrenal with a mechanism to regulate cortisol output according to exposure to hypoxia. One thing all three of these reviews show is that even after several decades of study into NO biosynthesis and function, there are clearly still many things left to discover.

scholarly journals Endothelial cell transfusion ameliorates endothelial dysfunction in 5/6 nephrectomized rats

2013 ◽  
Vol 305 (8) ◽  
pp. H1256-H1264 ◽  
Author(s):  
Maricica Pacurari ◽  
Dongqi Xing ◽  
Rob H. P. Hilgers ◽  
Yuan Yuan Guo ◽  
Zhengqin Yang ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Chronic Kidney Disease ◽  
Endothelial Dysfunction ◽  
Nitric Oxide Synthase ◽  
Kidney Disease ◽  
Endothelial Nitric Oxide Synthase ◽  
Vascular Function ◽  
Vascular Reactivity ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide

Endothelial dysfunction is prevalent in chronic kidney disease. This study tested the hypothesis that transfusion of rat aortic endothelial cells (ECs) ameliorates endothelial dysfunction in a rat model of chronic kidney disease. Male Sprague-Dawley rats underwent sham surgery or 5/6 nephrectomy (Nx). Five weeks after Nx, EC (1.5 × 106 cells/rat) or vehicle were transfused intravenously. One week later, vascular reactivity of mesenteric artery was assessed on a wire myograph. Sensitivity of endothelium-dependent relaxation to acetylcholine and maximum vasodilation were impaired by Nx and improved by EC transfusion. Using selective pharmacological nitric oxide synthase isoform inhibitors, we demonstrated that the negative effect of Nx on endothelial function and rescue by EC transfusion are, at least in part, endothelial nitric oxide synthase mediated. Plasma asymmetric dimethylarginine was increased by Nx and decreased by EC transfusion, whereas mRNA expression of dimethylarginine dimethylaminohydrolases 1 (DDAH1) was decreased by Nx and restored by EC transfusion. Immunohistochemical staining confirmed that local expression of DDAH1 is decreased by Nx and increased by EC transfusion. In conclusion, EC transfusion attenuates Nx-induced endothelium-dependent vascular dysfunction by regulating DDAH1 expression and enhancing endothelial nitric oxide synthase activity. These results suggest that EC-based therapy could provide a novel therapeutic strategy to improve vascular function in chronic kidney disease.

Fetal origins of adult vascular dysfunction in mice lacking endothelial nitric oxide synthase

2005 ◽  
Vol 288 (5) ◽  
pp. R1114-R1121 ◽  
Author(s):  
Monica Longo ◽  
Venu Jain ◽  
Yuri P. Vedernikov ◽  
Radek Bukowski ◽  
Robert E. Garfield ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Endothelial Nitric Oxide Synthase ◽  
Vascular Function ◽  
Vascular Dysfunction ◽  
Mesenteric Arteries ◽  
Fetal Origins ◽  
Uterine Environment ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide

Epidemiological studies have shown increased incidence of hypertension and coronary artery disease in growth-restricted fetuses during their adult life. A novel animal model was used to test the hypothesis regarding the role of an abnormal uterine environment in fetal programming of adult vascular dysfunction. Mice lacking a functional endothelial nitric oxide synthase (NOS3−/−KO, where KO is knockout) and wild-type (WT) mice (NOS3+/+WT) were crossbred to produce homozygous NOS3−/−KO, maternally derived heterozygous (NOS3+/−mat, mother with NOS3 deficiency), paternally derived heterozygous (NOS3+/−pat, normal mother), and NOS3+/+WT litters. Number of fetuses per litter was smaller in NOS3−/−KO and NOS3+/−mat compared with NOS3+/−pat and NOS3+/+WT mice. Adult female mice from these litters (7–8 wk old) were killed, and ring preparations of carotid and mesenteric arteries were mounted in a wire myograph to evaluate the passive and reactive vascular characteristics. Slope of the length-tension plot (a measure of vascular compliance) was increased, and optimal diameter (as calculated by Laplace equation) was decreased in NOS3−/−KO and NOS3+/−mat compared with NOS3+/−pat and NOS3+/+WT mice. Acetylcholine caused vasorelaxation in NOS3+/−pat and NOS3+/+WT and contraction in NOS3−/−KO and NOS3+/−mat mice. Responses to phenylephrine and Ca2+ were increased in NOS3−/−KO and NOS3+/−mat compared with NOS3+/−pat and NOS3+/+WT mice. Relaxation to isoproterenol was decreased in NOS3−/−KO and NOS3+/−mat vs. NOS3+/−pat and NOS3+/+WT mice. Abnormalities in the passive and reactive in vitro vascular properties seen in NOS+/−mat that developed in a NOS3-deficient maternal/uterine environment compared with the genetically identical NOS3+/−pat mice that developed in a normal environment are the first direct evidence in support of a role for uterine environment in determining vascular function in later life.

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